PMID- 37828896
OWN - NLM
STAT- MEDLINE
DCOM- 20231106
LR  - 20240210
IS  - 1754-8411 (Electronic)
IS  - 1754-8403 (Print)
IS  - 1754-8403 (Linking)
VI  - 16
IP  - 11
DP  - 2023 Nov 1
TI  - A method for TAT-Cre recombinase-mediated floxed allele modification in ex vivo 
      tissue slices.
LID - 10.1242/dmm.050267 [doi]
LID - dmm050267
AB  - Precision-cut lung slices (PCLS) are used for a variety of applications. However, 
      methods to manipulate genes in PCLS are currently limited. We developed a new 
      method, TAT-Cre recombinase-mediated floxed allele modification in tissue slices 
      (TReATS), to induce highly effective and temporally controlled gene deletion or 
      activation in ex vivo PCLS. Treatment of PCLS from Rosa26-flox-stop-flox-EYFP 
      mice with cell-permeant TAT-Cre recombinase induced ubiquitous EYFP protein 
      expression, indicating successful Cre-mediated excision of the upstream 
      loxP-flanked stop sequence. Quantitative real-time PCR confirmed induction of 
      EYFP. We successfully replicated the TReATS method in PCLS from Vangl2flox/flox 
      mice, leading to the deletion of loxP-flanked exon 4 of the Vangl2 gene. 
      Cre-treated Vangl2flox/flox PCLS exhibited cytoskeletal abnormalities, a known 
      phenotype caused by VANGL2 dysfunction. We report a new method that bypasses 
      conventional Cre-Lox breeding, allowing rapid and highly effective gene 
      manipulation in ex vivo tissue models.
CI  - (c) 2023. Published by The Company of Biologists Ltd.
FAU - Cheong, Sek-Shir
AU  - Cheong SS
AD  - National Heart and Lung Institute (NHLI), Imperial College London, London SW7 
      2AZ, UK.
FAU - Luis, Tiago C
AU  - Luis TC
AD  - Centre for Inflammatory Diseases, Department of Immunology and Inflammation, 
      Imperial College London, London W12 0NN, UK.
FAU - Stewart, Michelle
AU  - Stewart M
AD  - The Mary Lyon Centre at MRC Harwell, Harwell Campus, Oxfordshire OX11 0RD, UK.
FAU - Hillier, Rosie
AU  - Hillier R
AD  - The Mary Lyon Centre at MRC Harwell, Harwell Campus, Oxfordshire OX11 0RD, UK.
FAU - Hind, Matthew
AU  - Hind M
AUID- ORCID: 0000-0002-9966-9446
AD  - National Heart and Lung Institute (NHLI), Imperial College London, London SW7 
      2AZ, UK.
AD  - National Institute for Health Research (NIHR) Respiratory Biomedical Research 
      Unit at the Royal Brompton and Harefield NHS Foundation Trust, London SW3 6NP, 
      UK.
FAU - Dean, Charlotte H
AU  - Dean CH
AUID- ORCID: 0000-0002-8846-5472
AD  - National Heart and Lung Institute (NHLI), Imperial College London, London SW7 
      2AZ, UK.
LA  - eng
GR  - WT_/Wellcome Trust/United Kingdom
GR  - 210424/Z/18/Z/WT_/Wellcome Trust/United Kingdom
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20231103
PL  - England
TA  - Dis Model Mech
JT  - Disease models & mechanisms
JID - 101483332
RN  - EC 2.7.7.- (Cre recombinase)
RN  - EC 2.7.7.- (Integrases)
SB  - IM
MH  - Mice
MH  - Animals
MH  - Mice, Transgenic
MH  - Alleles
MH  - *Integrases/metabolism
MH  - Phenotype
PMC - PMC10629676
OTO - NOTNLM
OT  - Allele modification
OT  - Cell-permeant Cre recombinase
OT  - Gene manipulation
OT  - Precision-cut lung slices
OT  - Precision-cut tissue slices
OT  - TAT-Cre
COIS- Competing interests The authors declare no competing or financial interests.
EDAT- 2023/10/13 06:45
MHDA- 2023/11/06 06:42
PMCR- 2023/11/03
CRDT- 2023/10/13 03:43
PHST- 2023/04/24 00:00 [received]
PHST- 2023/10/06 00:00 [accepted]
PHST- 2023/11/06 06:42 [medline]
PHST- 2023/10/13 06:45 [pubmed]
PHST- 2023/10/13 03:43 [entrez]
PHST- 2023/11/03 00:00 [pmc-release]
AID - 333482 [pii]
AID - DMM050267 [pii]
AID - 10.1242/dmm.050267 [doi]
PST - ppublish
SO  - Dis Model Mech. 2023 Nov 1;16(11):dmm050267. doi: 10.1242/dmm.050267. Epub 2023 
      Nov 3.